The generally practiced method for the isolation of bacterial luciferase involving batch adsorption technique is described by J. Woodland Hastings, et.al., in Methods in Enzymology, Vol, LVII, 135 (1978) and A. Gunsalus-Miguel, et.al., J. Biol. Chem. 247(2), 398-404 (1972). Isolation of bacterial luciferase by affinity chromatography using immobilized flavin mononucleotide to bind the enzyme is described by C. A. Waters, et. al., in Biochem. Biophys. Res. Comm. 57, No. 4, 1152 (1974). Neither of these methods is particularly efficient in that each involves a considerable amount of time to perform and/or low yields of enzyme are recovered.
The method of isolating bacterial luciferase as described herein not only reduces significantly the amount of time required to isolate the enzyme, but also results in high yields of relatively pure enzyme.